1.0.

INTRODUCTION

The microbiological culture media was first studied at mid 19th century which is when the
bacteriologists attempted to grow microbes from using food or material which the microorganism had
been observed and also some compound. (Sandle, T. ,2011) The first one who had done the cultivation
of microorganism was Louis Pasteur, a chemist and microbiologist. He used ammonium salt, sugar, ash
and yeast as a media to cultivte microorganism. All of these meduim contained the basic need of
microbes to grow. (Tseng, C.K, 1946) Based on his obeservation, the types of chemical in medium can
lead to development of micrrorganisms and the copetition between microorganisms occur. Thus, there
is some species that outgrowing in culture rather than the other microbes.

After a few years passed, Robert Koch used liquid medium from solid media which is the broths
from fresh beef meat. He found out that this technique produced optimal growth for micoorganisms.
Koch, R, 1876) However, this technique has a disadvantage, which is the pathogen can recovered in the
medum. So, he improvised by adding gelatine in a meat. Then, the gelatine was replaced with agar and
the flat glass plate was replaced with petri dish to improvies the experiment. So, as we know, the culture
media found in either liquid state (broth) or in solid state (agar in petri dish).

In this experiment, the micrrorganisms are cultivated by using shake flask method. This method
nowadays has been widely used in culture research due to its simplicity, easy to handling and more
economically. (Büchs, 2001) The flask or know as Erlenmayer flask comes in different sizes and materials
and come with a choice wheter it is baffled or non-baffled. The medium in the flask is in the minimal
volume to maximize the gas transfer and it has baffle in the internal flask to promote efficient mixing
during shaken. The oxygen transfer rate in the flask mainly depend on the liquid volume and the
agitation speed and as the frequency of shaking increases, the more the effficiency of the oxygen
transfer. (S. Schiefelbein, 2013) During shaken, the changes in agitation velocity, the shaking diameter
and the size of flask will affect the hydrodynamics in the shake flask, thus will affect the growth of
microorganism. (O. Platas, 2013)
RECCOMMENDATION

 Gloves and mask must be wear when handling the bacteria e. coli to prevent from diseases such
as vomitting and diarrhoea. These pathogen might affect the health if it handle with carelessly.
 Alcohol must be sprayed at gloves consistantly to prevent the contamination at the media
culture during handling the broth.
 The equipment must be sterilised properly and must not be skipped to prevent contamination
as it can affect the growth of e. coli.
 Group members should take turns and take the reading of sample for every two hours and this
should be done in exact time to get the accurate results.
 The puvette should be clean in a proper way so that the absorbance value reading is accurate.

 Sandle, T. (2011): 'History and development of microbiological culture media', The Journal
(Institute of Science and Technology), Winter 2010-2011, pp10-14
 Tseng, C.K. (1946): Colloid Chemistry, Vol.6., in Alexander, J. (Ed.) Colloid Chemistry, New York:
Reinhold Publishing Corp., p629.
 Koch, R. (1876) Untersuchungen ueber Bakterien V. Die Aetiologie der Milzbrand-Krankheit,
begruendent auf die Entwicklungsgeschichte des Bacillus Anthracis. Beitr. z. Biol. D. Pflanzen 2:
277-310. In Milestones in Microbiology: 1556 to 1940, translated and edited by Thomas D.
Brock, ASM Press. 1998, p89
 Büchs, J. (2001). Introduction to advantages and problems of shaken cultures. Biochemical
Engineering Journal,7(2), 91-98. doi:10.1016/s1369-703x(00)00106-6
 Schiefelbein, S., Fröhlich, A., John, G. T., Beutler, F., Wittmann, C., & Becker, J. (2013). Erratum
to: Oxygen supply in disposable shake-flasks: Prediction of oxygen transfer rate, oxygen
saturation and maximum cell concentration during aerobic growth. Biotechnology Letters,35(8),
1231-1231. doi:10.1007/s10529-013-1237-z
 Platas, O. B., Sandig, V., Pörtner, R., & Zeng, A. (2013). Evaluation of process parameters in
shake flasks for mammalian cell culture. BMC Proceedings,7(Suppl 6). doi:10.1186/1753-6561-7-
s6-p17